Outlier of the whole study
We conducted a comprehensive analysis on gastritis and widely used TCM formulae and herbs for gastritis from the perspective of Cold/Hot ZHENG and network target in this study. First and foremost, based on seeds genes from Cold/Hot biological network [11] proposed by Li and microarray for CAG and CSG with Cold/Hot ZHENG [8], we constructed Cold/Hot ZHENG biological network for CG to discover the features and biomarkers of Cold/Hot ZHENG. On the basis of multi-omics data and machine learning algorithms, features and biomarkers of immune and metabolism were characterized and kept for following study. With the help of Cold/Hot biological network as well as the inferred immune and metabolism related characteristics, features and biomarkers of Cold/Hot ZHENG CG were acquired which were mainly related to immune regulation and metabolism.
Besides, we collected 29 formulae for GC and corresponding 132 herbs recorded in these formulae from Pharmacopoeia of China. Cold/Hot information and Meridian information of these herbs were collected from Pharmacopoeia. The distribution of Meridian information of the herbs were counted and the compounds composition of these herbs was obtained from commonly-used TCM database Herbiomap [12] and Symmap [13]. After filtering herbs without recording or without, 25 formulae, 89 Cold/Hot herbs, 19 other herbs and in total 2853 compounds were kept for further study, and the targets profiles for these herbs and formulae were characterized by our previous network-based algorithms [14, 15].
Finally, as a combination of Cold/Hot ZHENG biological network for CG and targets profiles for Cold/Hot herbs, a biological network describing the most-frequently targeted Cold/Hot genes of Cold/Hot herbs was constructed to uncover the mechanism of formulae with Cold/Hot properties and corresponding Cold/Hot herbs in formulae for CG to some extension. Based on the network and previous studies, potential biomarkers of Cold/Hot herbs in the formulae against CG were determined and their relationship with potential mechanism of Cold/Hot ZHENG were also explored to reveal the mechanism of these herbs against Cold/Hot ZHENG CG.
Molecular features of Cold/Hot ZHENG CG
In 2013, Li collected 35 patients of Cold/Hot ZHENG CG for microarray measurement, including 17 patients with Cold ZHENG (8 for chronic superficial gastritis and 9 for chronic atrophic gastritis) and 18 patients with Hot ZHENG (8 for chronic superficial gastritis and 10 for chronic atrophic gastritis). In order to find key molecules related to Cold/Hot ZHENG in CG, we collected seeds gene from previous Cold/Hot network model as background for Cold/Hot ZHENG. PLS-DA analysis successfully grouped Hot and Cold ZHENG CG for CAG patients and CSG patients, respectively (Fig. 1A). VIP (variable importance) for each gene in CAG patients or CSG patients was calculated and 26 of the seeds genes with VIP greater than 1 both occurred in CAG patients and CSG patients (Fig. 1B).
Analysis for finding representative molecules between Cold/Hot ZHENG CG. A PLS-DA analysis for Cold/Hot ZHENG CAG (left) and CSG (right). B Venn plot showing genes with VIP larger than 1 in both Cold/Hot ZHENG CAG and CSG. C Venn plot showing the differential expression of genes in microarray of Cold/Hot ZHENG CAG and CSG. D Heat map of 47 genes both up-regulated in Hot ZHENG CAG and CSG and 11 genes up-regulated in Cold ZHENG CAG and CSG for CAG patients (left) and CSG patients (right)
Besides, from another perspective, DEGs (differentially expressed genes) in CAG patients and CSG patients were calculated by limma model to find significantly expressed genes between Cold/Hot ZHENG in both CAG patients and CSG patients. Among these DEGs, 112 genes were differentially expressed in both CAG patients and CSG patients (adjust P value < 0.05, BH adjustment). And 11 genes were up-regulated in both Cold ZHENG CAG patients and CSG patients, while 47 of them were up-regulated in both Hot ZHENG of two disease conditions (Fig. 1C). Combining these two analyses of different perspectives, both in statistical methods and hierarchical clustering methods, Hot ZHENG patients were distinct from Cold ZHENG patients (Fig. 1D). This result suggested that there existed gene expression patterns between these two conditions of CG which might be able to mined from our analysis and further constructed biological networks.
Immune and metabolic characteristics of Cold/Hot ZHENG CG
Based on our found molecular features of Cold/Hot ZHENG CG, we further paid attention to the biological processes or pathways enriched by these molecular features. Firstly, we performed KEGG pathway enrichment [16] and Gene Ontology (GO) enrichment [17] on 1846 DEGs (1241 genes up-regulated in Cold ZHENG and 605 genes up-regulated in Hot ZHENG) of CAG patients. It was found that pathways and biological processes related to immune and metabolism were significantly enriched (Fig. 2A). Further, we performed Gene Set Enrichment Analysis (GSEA) [18] on CAG patient and GSEA terms that significantly enriched were shown in Fig. 2. It could be found that, GSEA terms related to immune, inflammation, cytokines and chemokines (Fig. 2B–D) were activated in Hot ZHENG CAG patient (also can be defined as inhibited terms in Cold ZHENG CAG patient, NES < − 1), while terms related to metabolism and secretion of peptide, hormone, steroid (Fig. 2B, E), as well as cellular junctions and adhesion were activated in Cold ZHENG CAG patient (NES > 1). These findings suggested that in Hot ZHENG CG, pathways or biological processes related to immune and inflammation might be over-developed, while in Cold ZHENG CG, the main distinguished features turned out to be activating in endocrine and energy metabolism. Apart from CG, these modules of immune regulation and metabolism had also been reported in researches about other diseases [19,20,21].
Enrichment analysis for immune and metabolic characteristics of Cold/Hot ZHENG CG A Dot plot of KEGG and GO enrichment analysis for 1846 DEGs of CAG patients. B GSEA for genes and their expression of CAG patients. C–E GSEA result for Chemokine Pathway, Inflammatory Response and Peptide Secretion, respectively
As inferred by our previous findings, we focused on the immune and inflammation characteristics in CG. Based on CIBERSORT algorithm [22], proportions of different immune cells were deconvoluted in Cold/Hot ZHENG CG, respectively. It could be found that some immune cells, represented by M1 macrophages, showed a significantly different proportion in Hot ZHENG CG than Cold ZHENG CG (Fig. 3A). The proportions of both M1 and M2 macrophages were significantly higher in Hot ZHENG CG than that of Cold ZHENG CG, which might confirm the findings that from the perspective of immune and inflammation regulation, the most distinguishing features between Hot ZHENG and Cold ZHENG gastritis is the over-inflammation in Hot ZHENG and the suppression of immune in Cold ZHENG.
Network construction of Cold/Hot ZHENG CG. A Inference of the proportion of immune cells significantly changed in Cold ZHENG CAG and Hot ZHENG CAG. B Expression of previous reported and newly found biomolecules for Cold/Hot ZHENG CAG in single-cell level of CAG patients. C Box plots showing the expression of biomolecules during the progression of gastric cancer
In addition, we filter out 3 Hot ZHENG CAG samples from large scale scRNA-seq of human gastric cancer progression. In this cellular level measurement of expression of genes in these characteristic pathways and biological processes related to immune regulation and metabolism, we focused on some key molecules in Cold/Hot ZHENG CG. It was found that biomarkers of Cold ZHENG CG which participated in these key pathways and biological processes, like HTR2B, CRH, NOS1 and LEP, hardly expressed in any cell types in Hot ZHENG CG samples. These genes were reported in previous study [8], and were found to be related to key link in Cold ZHENG, including 5-HT related gene HTR2B, corticotropin releasing hormone related genes CRH, CRHR1 and POMC, leptin related gene LEP and nitric oxide related gene NOS1. On the contrary, genes related to immune and inflammation were relatively higher expressed, especially in macrophages, which was consistent with our above findings that macrophages significantly increased in Hot ZHENG CG. These genes included CCL2, CD14, NFKB1, IL10RA, TNF and JAK2, which were related to inflammation, cytokines, chemokines and immune regulation (Fig. 3B). Based on public omics data, it was also found that some of these biomarkers showed significant changes in the progression from gastritis to dysplasia and gastric cancer (Fig. 3C), and were reported to participant in the progression of gastric cancer, such as LEP, CCL2, CD14 and TNF. The expression of these biomolecules and their related biomolecules was found to be associated with gastric cancer progression and prognosis [23,24,25,26], which also inferred that pathways or cells related to these biomarkers may also play roles in cancer progression and prognosis. These findings inferred us that the key biomolecules in Cold/Hot ZHENG might also play important roles in other disease progressions which need further analyses and researches. These complex features in immune regulation and metabolism, as well as biomolecules like TNF, VEGF, TGFB and NFKB1, also reflect the potential risk of CG, especially CAG in inflammation-induced tumorigenesis according to our previous constructed tumorigenesis network [27], and also be reported in researches of tumorigenesis in other digestive systems diseases like chronic hepatitis and enteritis [28, 29].
Finally, based on the combination of multi-omics data and machine learning algorithms, we constructed a homogeneous biological network, composed of key seeds genes of Cold/Hot ZHENG and DEGs in both two kinds of CG (Figure S1). In this network, the interactions between every two genes were collected from STRING database. It could be found that many of the seeds genes played important roles in these network with high connections, especially immune and inflammation-related genes like CCL2, CD14, NFKB1, IL2RB, JAK2, VEGFC, TGFB3 and IL10RA, most of which showed high expression in macrophages of Hot ZHENG CG patients. Besides, some genes related to endocrine and energy metabolism including SSTR2, SSTR5, HTR1A, CRH, CRHR1 and POMC also had high degrees in this network. It is worthy to be noticed that not only the biomolecules in the network for Cold/Hot ZHENG CG, but also the genes with close functional relationship or biological relationship may play important role in the further diagnosis and mechanism uncovering of Cold/Hot ZHENG CG.
According to our enrichment result, metabolism features of CG is also of vital importance in the diagnosis of Cold/Hot ZHENG CG. Metabolism related to peptide is significantly enriched, and the interactions between peptide and protein participate in various fundamental cellular functions [30]. The pathologically elevated steroid hormones may be accompanied by leptin resistance, which weakens normal energy expenditure and thermogenesis [31]. In our previous study, we found that serum level of leptin in CAG patients associated with Cold ZHENG was significantly higher than normal subjects [8]. Therefore, the presence of pathologically elevated leptin levels in patients with cold ZHENG means that their reduced energy expenditure and thermogenesis may be due to leptin resistance. Conditional Dlx1/2-null mice showed a loss of growth hormone-releasing hormone neurons with higher somatostatin expression and lower energy expenditure [32]. A previous study also showed that somatostatin in the paraventricular nucleus of the hypothalamus could inhibit thermogenesis [33], suggesting that SSTR involves in energy expenditure. It has been reported that 5-HT could inhibit thermogenesis through Htr3 in brown adipose tissue [34]. Besides, in the median preoptic nucleus, the thermoregulatory response is initiated by stimulation of GABA neurons, suggesting that GABA plays an important role in the process of immune regulation and energy expenditure [35]. Last but not the least, the suppression of tight junction and gap junction was associated with the activation of gene networks of adaptive immunity [36]. And tight junction was reported to be related to immune suppression in COVID-19 [37].
Characteristics of formulae for CG
From the perspective of system biology, we focused on the potential effect on formulae for CG. We measured some typical pathways or biological processes for Cold/Hot ZHENG CG in our above findings in all 29 formulae recorded in Pharmacopoeia. It was found that in specific pathways or biological processes in immune regulation, inflammation and steroid dominated energy metabolism, the potential effect of different formulae differed from others in immune-related pathways and biological processes like immune cells, immune response, cytokines and chemokine. Besides, some other pathways and biological processes related to energy reverse metabolism, nitric oxide. On the contrary, steroid metabolic process, response to steroid hormone, steroid hormone mediated signaling pathway, regulation of inflammatory response and response to oxidative stress were consistently significantly enriched and might be a coincident potential mechanism of these formulae against CG (Figure S2A).
All the 25 formulae could be pasted with six labels for traditional effects, including ZI YIN, XIAO JI, SAN HAN, QING RE, HUO XUE and XING QI. According to TCM experience, these six labels were corresponded with specific effects, in which ZI YIN, XIAO JI, SAN HAN, QING RE, HUO XUE and XING QI means nourishing humors, eliminating food stagnation, dispelling Cold, removing Hot, promoting the restoration of vitality and achieving smooth air flow, respectively. XING QI was the most frequent effect for these formulae, and it was positively correlated (Wilcoxon test, P value < 0.05) with the proportion of Hot herbs in a formula (Figure S2B, C). Besides, ZI YIN had significantly positive correlation with the proportion of Cold herbs, while SAN HAN had significantly positive correlation with the proportion of Hot herbs. In addition, XIAO JI and XING QI showed a relatively positively correlation with the proportion of Hot TCM in a formula. These findings were consistent with TCM theory that SAN HAN and XING QI are typical traditional effects of Hot and Warm herbs or formula with Warm and Hot properties, while QING RE and ZI YIN were the characteristics of Cold and Cool ones, and might uncover the material basis of these related six labels for traditional effects. Further, in the four significantly different labels for traditional effects in formulae with Cold/Hot herbs, we focused on the pathways of formulae with them to uncover the mechanism of these four traditional effects. It was found that pathways belonging to signal transduction, endocrine and immune were the most important in these four traditional effects, which might consistently support our findings that metabolism and immune regulation were the key mechanism of Cold/Hot ZHENG CG.
Herbs and depiction of their target profiles in formulae against CG
Another main finding of this study falls in the various mechanism of actions of Cold/Hot herbs in traditional formulae for CG. We collected 29 traditional formulae for CG and their corresponding herbs from Pharmacopoeia. These 29 formulae totally included 242 herbs (132 unique herbs). In annotating the compound composition for these herbs from two large databases Symmap and Herbiomap, 108 of these 132 herbs were successfully matched with the compounds as well as Cold/Hot information and meridian information. In total, 2853 unique compounds were found in these herbs and annotated with PubChem CID [38] for further targets prediction.
Target profiles of these compounds were calculated by our previous network-based algorithm DrugCIPHER-SC [14] and top 100 druggable targets of the profiles were chosen as the targets for further analysis. In order to measure the holistic targets of formulae and herbs, a previous statistical strategy [15] was performed and targets with occurrence significance less than 0.05 (BH adjustment) were chosen.
Cold/Hot information is one of the most important information in TCM herbs. According to Pharmacopoeia, the Hot and Cold properties have been divided into seven levels: Cold, light Cold, Cool, Ping (a kind of state which has no tendency to Cold or Hot), light Warm, Warm and Hot (including great Hot). The first three levels belong to Cold category and the last three levels belong to Hot category. Herbs belonging to Warm held the largest population with the amount of 45 and those of Cold held the second largest population of 31, while herbs belonging to Hot, light Warm and Cool held the least population.
Apart from Cold/Hot information, meridian was also of vital importance for herbs, for the reason that it might figure out the place where herbs took effect. Herbs in traditional formulae for CG mainly includes 12 kinds of meridian, including triple energizer (tri-jiao, a special TCM term), large intestine, small intestine, heart, pericardium, liver, lung, kidney, stomach, gallbladder, spleen and bladder. It was found that one herb might include more than one meridian, which showed that herbs might take effect in many tissues. It could be found that spleen was the most frequent destination of these herbs and liver, stomach, heart, lung and kidney ranked the second to the sixth. On the contrary, triple energizer, pericardium, gallbladder and bladder ranked at the end with the amount less than 10. Besides, we also observed the cross mapping of Cold/Hot properties and meridian (Fig. 4A), and it was worthy to be noticed that the most frequent pair was Warm and spleen. This finding was corresponding with previous hypothesis, since spleen is an important immune-related organ and one of the most important effect of Hot herbs is enhancing immune regulation. However, the distribution of Cold herbs seems to be more concentrated in stomach, kidney, liver, lung and heart rather than spleen.
Analysis of Cold/Hot herbs in formulae for CG. A Heat map showing the Cold/Hot properties and meridian of these Cold/Hot herbs. B, C Dot plot showing KEGG and GO enrichment of targets of Cold/Hot herbs. D Bar plot showing the proportions of the bidirectional regulatory effects of Cold/Hot TCM on representative targets
Molecular features of targets of cold/hot herbs for CG
To address the complexity of herbal compositions, many herbs may target the same biomolecules, as shown by network target analysis. To distinguish the tendencies of Cold and Hot herbs, we implemented a 0.7 threshold (refer to Methods) and categorized the targets into three groups: specific to Hot herbs, specific to Cold herbs, and shared by both. Shared targets are those potentially targeted by multiple Hot and Cold herbs simultaneously (Figure S3A). Cold herb targets include biomolecules such as TNF, ILR1, VEGFA, TLR2, and IL2RG, suggesting their roles in anti-inflammation and immune regulation (Figure S3C). Conversely, targets of Hot herbs are involved in energy metabolism processes including steroids, hormones, 5-HT, SSTR, NOS, CRH, and GABA, and key immune response molecules like IL6R, CXCR1 (Figure S3D). Additionally, shared targets involve key biological process participants like IL1B, CD4, AR, ESR1, ESR2, NFKB1, TGFB1.
KEGG and GO enrichment were also performed on Hot herbs targets and Cold herbs targets. It was found that, in the enrichment result of Cold herbs targets, biological processes related to inflammatory response, cytokines, chemokines and immune cells represented by macrophages were significantly enriched, as well as inflammatory pathways including TNF, VEGF, HIF-1a signaling pathways. Apart from these immune-related pathways or biological processes, those related to lipid metabolism were also significantly enriched, like fatty acid oxidation and lipid oxidation (Fig. 4B). The enrichment result of Hot herbs targets was much different, which mainly fell in biological processes bound up with inhibitory neurotransmitter like 5-HT, GABA, hormone including steroid hormone, corticosteroid hormone, peptide hormone and other endocrine hormones (Fig. 4C). Besides, the result also included cellular processes of T cells, like T cell activation and proliferation. In general, these findings showed that in the one hand, the mechanism of Cold herbs against Hot ZHENG mainly fell in immune-related and inflammation-related factors, as well as metabolism like lipid metabolism to some extent. On the other hand, the treatment of Hot herbs against Cold ZHENG CG were mainly related to neurotransmitter and endocrine, which further proved the closer relationship between Cold ZHENG CG and stress-induced factors, and showed therapeutic potential of Hot herbs in this regard.
Through public transcriptomics dataset of TCM intervened THP-1 cell lines in previous study [39], there were 21 Hot herbs and 17 cold herbs recorded in the dataset matched with the total 108 herbs for CG. Based on the transcriptomics data, it was possible for us to identify the regulatory direction of Cold/Hot herbs on these targets to some extent (Fig. 4D). It was found that targets like TNF, CCL2, CD86 and CSF1R showed consistently downregulation by both Cold/Hot herbs, and according to the literature, they played important roles in immune and inflammation regulation [40,41,42,43]. On the other hand, targets like HSPA1A and MAPK10 were both up-regulated by Cold/Hot herbs. Besides, targets like TGFB1, ICAM1 received up-regulation and down-regulation from many Cold/Hot herbs while IL6 and VEGFA were subject to bidirectional regulation but primarily downregulated by both Cold/Hot herbs. In additional, SSTR2 were found to be decreased by Cold/Hot herbs (Figure S3E), which might lead to the change of Somatostatin (SS) [44, 45]. It was worth to be noticed that SS and its receptor played important roles in gastric diseases [46,47,48], and its absence might induce GC [49].
Bidirectional regulatory effect of TCM in cold and hot CG
These results preliminarily revealed the mechanisms of action of Cold/Hot herbs in the treatment of CG associated with Cold/Hot ZHENG (Fig. 5). These functional characteristics of Cold/Hot herbs in CG formulae for immune regulation and metabolism regulation were also found to be therapeutic targets in researches of other formulae for other diseases [50,51,52,53].
Tai Chi Diagram showing the regulation programs of Cold/Hot herbs dominated by bi-directional regulation on inflammation/immune and energy metabolism. Cold herbs showed a trend to suppress the over-inflammation and over exuberant energy metabolism in Hot ZHENG, while Hot herbs preferred to enhance and recover immunity and energy metabolism in Cold ZHENG
Based on what we found for molecular features of Cold/Hot herbs, two specific networks for the Cold/Hot herbs targets respectively, according to the interaction recoded in STRING database (Figure S4). In Cold herbs targets, INS, TLR2, VEGFA, TNF, IL1R1 and TGFB3 showed a vital role in the targets network of Cold herbs for CG. And in Hot herbs targets, 5-HT related genes HTR1A and HTR1B, NOS1, GABA-related genes GAB family and CRH, as well as somatostatin receptor including SSTR2 and SSTR5 were found in the targets network of Hot herbs for CG. In other words, Hot herbs might regulate Cold ZHENG gastritis through regulating endocrine and energy metabolism. Apart from these genes, targets like CCL2, IL6, JAK2, IL2RA and CXCR1 were also of vital importance in the Hot herbs’ targets network, which might infer us that another potential mechanism of Hot herbs against Cold ZEHNG gastritis was to regulate immune responses.
Immune-related pathways and biomolecules like TLR2 and CD14 were potential targets of Weifuchun capsule, which was clinically used for CAG, composed of Cold/Hot herbs and performed effect on both Cold/Hot ZHENG, like regulating immune response and anti-inflammation [54]. Huangqi Jianzhong decoction, a formula for Cold ZHENG CG, showed protective effects in CAG rats might be due to the balance of energy expenditure [55]. Taken together, the thermogenesis and immune-enhancing effects of Hot herbs may contribute to the therapeutic effect on Cold ZHENG CG. For herbs with cold property, their treatment of Hot ZHENG CG relies mainly on their anti-inflammatory effects, such as involving the TNF signaling pathway, NF-κB signaling pathway, and VEGF signaling pathway. Zuojin Pill is used in the treatment of Hot CG because it contains the cold property herb (Huanglian, dried rhizome of Coptis chinensis Franch., Coptis teeta Wall., and Coptis deltoidea C.Y.Cheng and P.K.Hsiao), which has been shown to have anti-inflammatory effects on CAG in rats by inhibiting the NF-κB signaling pathway [56]. Another widely used formula for CAG, Moluodan, was reported to reduce the inflammation level, as well as increasing lipid accumulation in MNNG-induced cells [57]. Serum levels of TNF-α, IL-8, and VEGF have been reported to be associated with the severity of CG, the severe degree of neutrophil infiltration in CG, and the severity of precancerous lesions in the stomach, respectively [58]. Weiqi decoction formula has been reported to reduce VEGF levels in CAG rats [59]. The inhibition of the inflammatory response is the main therapeutic route for cold property herbs in the treatment of Hot ZHENG CG. Besides, leptin is considered to be a link between the neuroendocrine and immune systems and could be a possible target for intervention in immunometabolism-mediated pathophysiology and it has been reported that leptin resistance individuals have lower NK cell count and function than normal individuals [60]. Gastric mucosal leptin expression was significantly higher in H. pylori-positive patients than in negative patients [61]. Thus, in the context of immune-metabolic imbalances, leptin appears to be the pivotal molecule in the treatment of Cold/Hot CG with Cold/Hot herbs.
Transcriptomic level validation of the cold/hot regulatory effects of TCM
To demonstrate the role of TCM formulae in treating gastritis from the perspective of Cold and Hot, we selected a classic formula, Jin Hong tablets (a formula contains one Cold herb, Toosendan Fructus and three Warm herbs), which might have bi-directional regulatory effects from the perspective of Cold/Hot, constructed gastritis models and validated our findings through transcriptomics and molecular detection. Based on the differentially expressed genes identified in blood samples and gastric tissue samples (Fig. 6A, B), KEGG pathway enrichment analysis was performed on upregulated and downregulated genes (Fig. 6C). It was found that pathways related to Organismal systems, especially immune systems and endocrine system were significantly changed after the intervention of Jin Hong tablets. For example, estrogen signaling pathway were significantly enriched (Fig. 6D), and was reported to be of vital importance in both energy metabolism [62,63,64] and immune regulation [65, 66], which was consistent with our findings. More specifically, we observed the enrichment of differentially expressed genes in biological processes (Fig. 6E) and it was found that in gastric tissues, a significant number of metabolic-related biological processes underwent notable changes, including those related to peptides, glucose, and others. On the other hand, in blood, a significant number of immune-related biological processes were altered, including those related to immune response, cytokines, and various immune cells.
Experiments validation from the perspective of Cold/Hot ZHENG. A–B Volcano plots for differential expressed genes before and after the intervention of Jin Hong tablets in blood and stomach, respectively. C Dot plot showing pathways with enrichment of genes differential expressed in blood and stomach, as well as their classification (dots below 0 were enriched by downregulated genes and dots above 0 were enriched by upregulated genes). D Representative enrichment of GSEA. E Enrichment result of the siginificantly changed pathways or biological processes in blood and stomach before and after the intervention of Jin Hong tablets. F Bar plot showing the proportions of immune cells in different samples before and after the intervention of Jin Hong tablets. G Inferred proportions of two types of T cells before and after the intervention of Jin Hong tablets. H Bar plot showing the expressions of IL-2, IL-6, SS and Leptin in blood
By deconvolving the transcriptomic expression matrix, we depicted the proportion changes of 6 major immune cells in gastric tissues before and after intervention, as well as in blood before and after intervention (Fig. 6F). For example, the proportions in CD8 + T cell and regulatory T cell undergone significantly changed after the intervention of Jin Hong tablets (Fig. 6G), which to some extent represented the restoration of immune response capabilities [67]. Finally, we measured the core targets in Cold/Hot biological networks, including IL-2, IL-6, SS and Leptin and found that their expression significantly recovered to normal levels (Fig. 6H).
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